GENERAL PROPERTIES OF VIRUSES

Definition:-
Viruses are the smallest infectious agents
(20-300 nm in diameter), containing one kind
of nucleic acid (RNA or DNA) in their
genome. The nucleic acid is encased in a
protein coat, which may be
surrounded by a lipid envelope.
The entire infectious unit
is termed a virion.
Viruses are inert in the extracellular

environment; they replicate only in living

cells, being obligatory intracellular

parasites.

Viruses can infect man, animals, bacteria,

insects and plants.

Classification:-
Most of the clinically important viral pathogens
can be categorized into groups according to
their structural characters into:
1- RNA non enveloped (naked) viruses.
2- RNA enveloped viruses.
3- DNA non enveloped (naked) viruses.
4- DNA enveloped viruses.

However, some viruses e.g. tumor viruses and

slow viruses are best described in terms of their
biologic features.
 CLASSIFICATION OF VIRUSES

Non-enveloped enveloped

1
2
• All DNA viruses are unenveloped (naked)except 1
• All DNA viruses are double stranded except 2 1
• All DNA viruses replicate in the nucleus except poxvirus
 CLASSIFICATION OF VIRUSES

• All RNA viruses are enveloped except

- Picorna viruses
- Reoviruses
❑ All RNA viruses are single stranded (ss) except reoviruses
❑ All RNA viruses replicate in the cytoplasm except retrovirus and
influenza virus
Size and Shape:-
Viruses range from 20-300nm (nm = 1/1000
um) in diameter. Their shapes are frequently
referred to as spheres, bullets, rods or bricks,
but in reality they are complex structure of
precise geometric symmetry. The shape of
virus particles is determined by the
arrangement of the repeating subunits that
form the protein coat (capsid) of the virus.
 The size of many viruses
has been determined by
********
1- Direct observation in the electron
microscope.
2- Filtration through membranes of graded
porosity.
3- Sedimentation in the ultracentrifuge.
4- Comparative measurements to other
agents.
Viral Structure:
The virus particle (virion), consists of a nucleic
acid core called “virus genome” surrounded
by a protein coat called the capsid.
Some viruses are enveloped, while others are
naked.
Some viruses may contain glycoprotein spikes.
Viral Nucleic Acids:-
The viral nucleic acid is located internally and
can be either single or double stranded either
DNA or RNA.
The nucleic acid can be either linear or circular.
The DNA is always a single molecule; the RNA
can exist as a single molecule or in several
pieces (segmented).
Capsid and Symmetry:-
The viral protein coat (capsid) consists of
repeated subunits called capsomers. The
arrangement of capsomers gives the virus
structure its geometric symmetry.
There are three forms of symmetry.
1- Icosahedral
2- Helical
3- Complex structure
1- Icosahedral; in which the capsomers are
arranged in 20 triangles forming a symmetric
figure (an icosahedron) e.g adenoviruses.

2- Helical; in which the capsomers are

arranged in a hollow coil that appears
as a spiral or a helix around a spiral
nucleic acid e.g. orthomyxoviruses.

3- Complex structure; some viruses are

Complicated in structure e.g. bacterio-
phages have spermatozoa like structure.
Viral Proteins:-
Are either outer capsid proteins or internal proteins.
They have several important functions:
1- The Outer Capsid Proteins
a- They protect the genetic material.
b- They mediate the attachment of the virus to specific
receptors on the host cell surface.
c- They are important antigens that induce neutralizing
antibody and activate cytotoxicTcells.
d- They determine the type specificity of the virus i.e.
poliovirus types 1,2 and 3 are distinguished by antigens of
their capsid proteins. It is important to know different
serotypes of a virus in vaccine preparations.
e- Some surface proteins e.g. influenza virus hemagglutinin,
cause agglutination of red blood cells.
2- Internal Proteins:
a - Some of the internal proteins are
associated with the nucleic acid e.g.
polymerases and reverse transcriptase
(essential for virus replication).
b - Some viruses produce proteins that act
as superantigens e.g. Epstein-Barr
virus and cytomegalovirus.
Envelope:-
The envelope is a lipoprotein membrane composed of
lipid derived from the host cell membrane and a
protein that is virus- specific.
Furthermore, there are frequently glycoproteins in
the form of spike like projections on the surface
which attach to host cell receptors during the entry
of the virus into the cell.
Another protein, matrix protein lies between the
capsid proteins and the envelope.
It was noticed that the presence of an envelope
confers instability to the virus. Enveloped viruses
are more sensitive to heat, detergents and lipid
solvents such as alcohol and ether than are non
enveloped (naked) viruses.
Atypical Virus Like Agents:-
1- Defective viruses: are composed of viral nucleic
acid and proteins, but cannot replicate without a
helper virus that provides the missing function.
2- Pseudovirions: contain host cell DNA instead of
viral DNA within the capsid. They can infect cells
but can not replicate.
3- Viroids: consist solely of a single molecule of RNA
without a protein coat or envelope. They cause
several plant diseases.
4- Prions: are infectious particles that are composed
solely of protein i.e. they contain no detectable
nucleic acid. They are implicated as the cause of
certain slow diseases as bovine spongiform
encephalopathy, known as mad cow disease.
Prions:
Are remarkably resistant to heat, ultraviolet light or
formaldehyde, however they are inactivated by
strong detergent agents and autoclaving.
Because prions are resistant to the temperature
of cooking, it is suspected that they are transmitted
by food.
CULTIVATION OF VIRUSES:-
Culture of viruses requires living cells. These include:
a) Embryonated chicken eggs: Inoculation of
chorioallantoic membrane, amniotic sac, the
allantoic sac, the yolk sac or the embryo itself
may be used.
b) Animal inoculation:
It is used mainly to study pathogenesis of
viral diseases, biologic assays and virus
oncogenesis.
Embryonated eggs are better than animal
inoculation because:-
-They are sterile.
-They have no immunological function.
-They are easily available and inexpensive.
c) Tissue culture:
This is the most widely used.

Three types of cell lines are in common use:

- Primary cell lines: prepared from organ fragments

e.g monkey kidney. Such cells

can only divide for several

passages (5) then die.

- Diploid cell lines: prepared from human
embryonic fibroblasts. They grow rapidly and
can divide up to 50 passages.

Continuous cell lines:

are cells capable of indefinite
growth and derived from
malignant cells e.g. human
carcinoma of the cervix
e.g. Hela cells.
Detection of Virus Infected Cells:
The viral growth is detected in tissue culture by:
1- Cytopathic effect (CPE).
2- Plaque formation.
3- Inclusion bodies.
4- Detection of virus antigens.
5- Hemadsorption.
6- Neutralization test.
7- Detection of viral nucleic acid.
1- Cytopathic effect (CPE): may be in the form of:
a- Cell lysis or necrosis.
b- Production of multinucleated giant cells.

2- Plaque formation: Plaques are virally infected

areas in the cell line. They can be
seen by the naked eye after
staining with vital stains.
Rt bottle of uninfected tissue culture monolayer.

Lf bottle of infected tissue culture showing plaques.

3- Inclusion bodies: In the course of virus replication
, virus- specific structures may be produced. These
structures are called “inclusion bodies”. These
may be intranuclear, or intracytoplasmic or in both
nucleus and cytoplasm. Several names are given
to the inclusion bodies and their presence inside
the cell is diagnostic, e.g. Negri bodies in rabies
rabies.
Negri bodies
4- Detection of virus antigens; released in the
tissue culture medium by different immunologic
assays e.g. by ELISA, hemagglutination.
5- Hemadsorption; clumping of RBCs when added
to cells infected with hemagglutinating viruses
e.g. influenza virus.
6- Neutralization test: Neutralization of the CPE of a
virus on tissue culture by specific antisera.
7- Detection of viral nucleic acid by molecular-
based assays such as PCR. They provide rapid
and sensitive methods of detection.
VIRUS REPLICATION:-
Viruses multiply only in living cells. The host cell
provides the energy and low molecular weight
precursors for synthesis of viral proteins and nucleic
acids.
General Steps in Viral Replication Cycles:-
1- attachment.
2- Penetration.
3- Uncoating.
4- Eclipse.
5- Intracellular synthesis.
6- Assembly and release.
1- Attachment:
Virus replication starts with the interaction of
a virion with a specific receptor site on the
surface of a host cell. These receptors are
glycoprotein. Each susceptible cell may
contain up to 100,000 receptor sites for a
given virus.

2- Penetration:
After binding, virus particle is either taken up
inside the cell by a process of endocytosis
within endosomes or directly penetrate
across the plasma membrane.
3- Uncoating:
It occurs shortly after penetration of the
virus. It is a physical separation of the viral
nucleic acid from its outer components.
The genome may be released as free
nucleic acid or as nucleocapsid. The
nucleocapsid usually contains viral
polymerases, essential for synthesis of
viral components. Uncoating requires
some cellular enzymes.
4- Eclipse:
Is the period after penetration during which no
infectious virus components can be detected
inside the host cell. During this phase, the cell is
redirected toward synthesizing early proteins
(enzymes) which are essential for viral replication.

5- Intracellular synthesis:
The first step is mRNA synthesis. Once viral
mRNA is synthesized, it is translated by host cell
ribosomes and tRNA into viral proteins which are
early proteins and late proteins (structural
proteins).
 Replication of viral genome depends on
complementarity i.e. a complementary
strand to the viral genome is synthesized,
this strand then serves as the template for
the synthesis of the actual viral genome
(enzymes required for replication of viral
genome) .
5- Assembly and Release:
The progeny particles are assembled by
packaging the viral nucleic acid within the
capsid protein. Virus particles are released
from the cell by either of two processes:-
a- Rupture of the cell membrane and
release of the unenveloped particles.
b- Release of enveloped viruses by budding
through the outer cell membrane.
 PATHOGENESIS OF VIRUS INFECTIONS
Viruses should reach a susceptible cell before they
can produce disease. Therefore, they should have:
1- Portal of entry; namely the respiratory tract, gastro-
intestinal tract skin, urogenital tract or conjunctiva.
2- A pathway through the body; namely the blood, the
lymphatics or the nerves.
3- A target organ which may be CNS, skin, liver..
Viruses may produce local or systemic infections
which may reach the full blown picture or more
commonly end in a subclinical form. Some viruses
persist.
I- Local infections :
Infection occur at the portal of entry with no viraemia :
- Influenza and common cold at the mucous
membrane of the respiratory tract.
- Rotavirus infection at the GIT causing diarrhoea.
- Papilloma virus infection of the skin causing human
warts.
Local infections are characterized by:
1- Short incubation period.
2- Short lasting immunity mediated by IgA and
interferon.
II- Systemic infections:
After primary replication at the site of entry , the virus
travels through the blood or lymphatics causing
viraemia, or through the nerves to reach a target
organ that has specific receptors for the virus.
Systemic infections are characterized by:
1- Long incubation period.
2- Long lasting immunity mediated by IgG and IgM.
3- Infection can be stopped at the viraemic stage by
immune mechanisms (neutralizing antibodies).
This leads to subclinical or abortive infection.
4- Gamma globulins given to contacts of a case may
abort infection if given during the incubation
period before the viraemic stage.
Ill- Persistent viral infections:
Sometimes viruses persist for a long time in the host
in one of the following forms:
a- Chronic infections in which the virus can be
continuously detected with no or mild symptoms
e.g. hepatitis B chronic carriers.
b- Latent infections in which the virus persists
hidden most of the time, with periodic
reactivation and development of clinical
lesions containing the virus e.g. herpes
simplex virus infections.
c- ”SIow virus” infections: These have a very long
incubation period of months or years with no
clinical symptoms e.g. HIV and other prions as
mad cow disease
 Laboratory diagnosis of viral
infections:
A. Direct detection of viruses, their antigens, or
their nucleic acids in clinical specimens.
1)Ultrastructural studies:
• physical measurements of virus
particles:Determinations of their size by
filteration through colloidal membranes of
various pore sizes
• chemical investigation:Determine the overall
composition of viruses
2)light microscopy :

This can be used to visualize large viruses

e.g. poxviruses. Inclusion bodies can be
seen in several viral infections. In rabies
,intracytoplasmic inclusion bodies called
negri bodies can be detected in nerve
cells.
3)electron microscopy:

It provides the following information

The absolute number of virus paticles

present in any preparation(total count)

The appearance\ structure of the particles.

4)immunoelectron microscopy(IEM):

addition of specific antisera to the clinical

material leads to aggregation of virus, so it
can be detected more readily than
separate virus particles e.g. diagnosis of
hepatitis A virus and rotavirus in stools.
5)immunofluorescence microscopy:
detection of virus in smears from lesions
using fluorescein labeled specific antisera.

6)solid-phase immunoassays:

both RIA and ELISA can be used for

detection of viral antigens in different
clinical specimens e.g. detection of
hepatitis B surface antigen in blood.
7)nucleic acid hybridization:using DNA
probes it is possible to detect virus nucleic
acid in clinical specimens or tissue
samples.
8)polymerase chain reaction(PCR):this
technique involves amplification of a short
sequence of a target DNAor RNA leading
to accumulation of large amounts of that
sequence, so it can be easily detected
.PCR can be used to determine the
quantity of viruses in patient blood (virus
load)as in HIV to monitor the course of the
disease.
B.Isolation of viruses

• Cell culture methods

• Animal inoculation

• Chick embryo
C.Serological detection of antiviral
antibodies:
• The antibody response to a viral infection
normally starts with the relatively transient
production of IGm followed by a long
lasting production of IGg . So a specific
IGM can be taken as a sign of recent
infection.
• Various serological methods for viruses
include:
• Haemagglutination inhibition tests of
red blood cells e.g. influenza virus.

• Complement fixation tests.

• Radioimmunoassay.

• Immunofluorescence.

• ELISA.

• Western blot assay.

• Monoclonal antibodies:
• This enabled virologists to look not only at
the whole virus, but at specific regions-
epitope- of individual virus antigens.
 Antiviral Chemotherapy

• The few antiviral drugs used act by

stopping virus attachment,
penetration, uncoating, or intracellular
synthesis.
 Classes of antiviral agents :

A-natural antiviral compound:

Interferon is natural substance that has
antiviral properties in adjacent,
noninfected cells.
 Types of interferons:

1)interferon-α(IFN-α) has maximal antiviral

activity.

2)Interferon-β(IFN-β) has intermediate

antiviral activity.

3) Interferon-γ(IFN- γ )is more active as a

lymphokine than as antiviral agent.
 B-Synthetic antiviral agents

antiviral compounds can be grouped

according to their point of action in the
viral replication cycle.

1-nucleoside analogs:

they block viral nucleic acid synthesis e.g.

acyclovir,ribavirin and zidovudine(AZT).
1- Azidothymidine (AZT. Zidovudine):
- Synthetic thymidine analogue.
- Inhibits replication of HIV by inhibiting viral
reverse transcriptase and blocking proviral
DNA synthesis.
- Reduces morbidity and mortality in AIDS
patients
- Expensive. causes bone marrow
depression
• Dideoxyinosine (ddi): Has same actions
as AZT but less toxic

2- Deoxythiacytidine and Stavudine:

Nucleoside analogues used if resistance
to treatment with AZT or ddi.
3- Acyclovir:

- Guanosine analogue inhibiting Herpes

simplex virus.

- Inhibits virus specific DNA polymerase.

- Used - topically in treating herpetic corneal

ulcers and skin lesions

- Parenterally in serious systemic infections:

herpes encephalitis.
Analogs Block DNA Synthesis

Figure 20.16b, c
2- nucleotide analogs:
they differ from the nucleoside analogs in
having an attached phosphate group.
Their ability to persist in cells for long
periods of time increases their potency
e.g. cidofovir.
3-non nucleoside reverse
transcriptase inhibitors:
They act by binding directly to reverse
transcriptase and disrupt the enzyme
catalytic site e.g. Nevirapine.
4- protease inhibitor:
such drugs inhibit the viral protease that is
required at the late stage of replicative
cycle. Inhibition of protease yields non
infectious virus particles e.g. Saquinavir
used for treatment of HIV infection.
 5- other types antiviral agents:

-Ribavirin:
- Synthetic nucleotide effective against many
DNA & RNA viruses

-Acts by interfering with mRNA synthesis

- Amantadine & Rimantadine:
- synthetic amines inhibiting uncoating of influenza A but
not B virus.

- Used prophylactically against influenza A

• Zanamivir inhibits the release of influenza virus from

infected cells

• foscarnet: it is a viral polymerase inhibitor of herpes

viruses. It also inhibits the reverse transcriptase of
HIV.